Peroxiredoxin promotes longevity and H2O2-resistance in yeast through redox-modulation of protein kinase A

AbstractPeroxiredoxins are H2O2scavenging enzymes that also carry H2O2signaling and chaperone functions. In yeast, the major cytosolic peroxiredoxin, Tsa1 is required for both promoting resistance to H2O2and extending lifespan upon caloric restriction. We show here that Tsa1 effects both these functions not by scavenging H2O2, but by repressing the nutrient signaling Ras-cAMP-PKA pathway at the level of the protein kinase A (PKA) enzyme. Tsa1 stimulates sulfenylation of cysteines in the PKA catalytic subunit by H2O2and a significant proportion of the catalytic subunits are glutathionylated on two cysteine residues. Redox modification of the conserved Cys243 inhibits the phosphorylation of a conserved Thr241 in the kinase activation loop and enzyme activity, and preventing Thr241 phosphorylation can overcome the H2O2sensitivity of Tsa1-deficient cells. Results support a model of aging where nutrient signaling pathways constitute hubs integrating information from multiple aging-related conduits, including a peroxiredoxin-dependent response to H2O2.

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Peroxiredoxin promotes longevity and H2O2-resistance in yeast through redox-modulation of protein kinase A

eLife ◽

10.7554/elife.60346 ◽

2020 ◽

Vol 9 ◽

Cited By ~ 2

Author(s):

Friederike Roger ◽

Cecilia Picazo ◽

Wolfgang Reiter ◽

Marouane Libiad ◽

Chikako Asami ◽

...

Keyword(s):

Protein Kinase ◽

Protein Kinase A ◽

Significant Proportion ◽

Catalytic Subunits ◽

Kinase Activation ◽

H2o2 Resistance ◽

Nutrient Signaling ◽

Scavenging Enzymes ◽

Pka Pathway ◽

Kinase A

Peroxiredoxins are H2O2 scavenging enzymes that also carry out H2O2 signaling and chaperone functions. In yeast, the major cytosolic peroxiredoxin, Tsa1 is required for both promoting resistance to H2O2 and extending lifespan upon caloric restriction. We show here that Tsa1 effects both these functions not by scavenging H2O2, but by repressing the nutrient signaling Ras-cAMP-PKA pathway at the level of the protein kinase A (PKA) enzyme. Tsa1 stimulates sulfenylation of cysteines in the PKA catalytic subunit by H2O2 and a significant proportion of the catalytic subunits are glutathionylated on two cysteine residues. Redox modification of the conserved Cys243 inhibits the phosphorylation of a conserved Thr241 in the kinase activation loop and enzyme activity, and preventing Thr241 phosphorylation can overcome the H2O2 sensitivity of Tsa1-deficient cells. Results support a model of aging where nutrient signaling pathways constitute hubs integrating information from multiple aging-related conduits, including a peroxiredoxin-dependent response to H2O2.

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Identification of a clinically homogenous subgroup of benign cortisol-secreting adrenocortical tumors characterized by alterations of the protein kinase A (PKA) subunits and high PKA activity.

Acta Endocrinologica ◽

10.1530/eje-07-0819 ◽

2008 ◽

Vol 158 (6) ◽

pp. 829-839 ◽

Cited By ~ 17

Author(s):

C Vincent-Dejean ◽

L Cazabat ◽

L Groussin ◽

K Perlemoine ◽

G Fumey ◽

...

Keyword(s):

Protein Kinase ◽

Protein Kinase A ◽

Carney Complex ◽

Catalytic Subunits ◽

Regulatory Subunits ◽

Adrenocortical Tumors ◽

Camp Pathway ◽

Adrenocortical Adenomas ◽

Pka Pathway ◽

Kinase A

ObjectiveThe cAMP/protein kinase A (PKA) pathway plays an important role in endocrine tumorigenesis. PKA is a heterotetramer with two regulatory subunits (four genes:PRKAR1A,PRKAR1B,PRKAR2A,PRKAR2B) and two catalytic subunits. InactivatingPRKAR1Amutations have been observed in Carney complex and a subset of adrenocortical tumors (ACT). This study was designed to search for other alterations of PKA in ACT, and to establish their correlation with the clinical characteristics.MethodsIn this study, 35 ACT (10 non-secreting adrenocortical adenomas (ACA-NS), 13 cortisol-secreting adenomas (ACA-S), and 12 malignant s (ACC)) were studied. PKA subunits were studied by western blot and RT-qPCR. The PKA activity was measured.ResultsA subgroup of ACA-S with a 96% R2B protein decrease by comparison with normal adrenal (4.1%±4 vs 100%±19,P<0.001) was identified, ACA-S2 (6/13). By contrast, no differences were observed in ACC and ACA-NS. The level of R1A mRNA was decreased in ACA-S (P<0.001), but not the level of R2B mRNA. No mutation of the R2B gene was detected in ACA-S2. The ACA-S2 group with loss of R2B protein showed a threefold higher basal PKA activity than the ACA with normal R2B protein (3.37±0.31 vs 1.00±0.20,P<0.0001). The ACA-S2 tumors with the loss of the R2B protein presented a homogenous phenotype and were all small benign cortisol-secreting tumors.ConclusionThis loss of PRKAR2B protein due to a post-transcriptional mechanism in ACA-S is a new mechanism of cAMP pathway dysregulation in adrenocortical tumorigenesis. It defines a new subtype of secreting adenomas with high basal PKA activity presenting a homogenous clinical phenotype.

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Decision letter: Peroxiredoxin promotes longevity and H2O2-resistance in yeast through redox-modulation of protein kinase A

10.7554/elife.60346.sa1 ◽

2020 ◽

Keyword(s):

Protein Kinase ◽

Protein Kinase A ◽

Redox Modulation ◽

H2o2 Resistance ◽

Kinase A

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Cooperative Activation of Lipolysis by Protein Kinase A and Protein Kinase C Pathways in 3T3-L1 Adipocytes

Endocrinology ◽

10.1210/en.2004-0803 ◽

2004 ◽

Vol 145 (11) ◽

pp. 4940-4947 ◽

Cited By ~ 29

Author(s):

Katrin Fricke ◽

Aleksandra Heitland ◽

Erik Maronde

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Protein Kinase A ◽

Signaling Pathways ◽

Hormone Sensitive Lipase ◽

Glycerol Release ◽

Kinase C ◽

Lipolytic Effect ◽

Pka Pathway ◽

Kinase A

Abstract In the present study, we investigate the coherence of signaling pathways leading to lipolysis in 3T3-L1 adipocytes. We observe two linear signaling pathways: one well known, acting via cAMP and protein kinase A (PKA) activation, and a second one induced by phorbol 12-myristate 13-acetate treatment involving protein kinase C (PKC) and MAPK. We demonstrate that both the PKA regulatory subunits RIα and RIIβ are expressed in 3T3-L1 adipocytes and are responsible for the lipolytic effect mediated via the cAMP/PKA pathway. Inhibition of the PKA pathway by the selective PKA inhibitor Rp-8-CPT-cAMPS does not impair lipolysis induced by PKC activation, and neither PD98059 nor U0126, as known MAPK kinase inhibitors, changes the level of glycerol release caused by PKA activation, indicating no cross-talk between these two pathways when only one is activated. However, when both are activated, they act synergistically on glycerol release. Additional experiments focusing on this synergy show no involvement of MAPK phosphorylation and cAMP formation. Phosphorylation of hormone-sensitive lipase is similar upon stimulation of either pathway, but we demonstrate a difference in the ability of both PKA and the PKC pathway activation to phosphorylate perilipin, which in turn may be an explanation for the different maximal lipolytic effect of both pathways.

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B-Raf-dependent regulation of the MEK-1/mitogen-activated protein kinase pathway in PC12 cells and regulation by cyclic AMP

Molecular and Cellular Biology ◽

10.1128/mcb.14.10.6522-6530.1994 ◽

1994 ◽

Vol 14 (10) ◽

pp. 6522-6530

Author(s):

R R Vaillancourt ◽

A M Gardner ◽

G L Johnson

Keyword(s):

Growth Factor ◽

Protein Kinase ◽

Map Kinase ◽

Protein Kinase A ◽

Pc12 Cells ◽

Kinase Activation ◽

Mitogen Activated Protein ◽

Epidermal Growth ◽

Camp Levels ◽

Kinase A

Growth factor receptor tyrosine kinase regulation of the sequential phosphorylation reactions leading to mitogen-activated protein (MAP) kinase activation in PC12 cells has been investigated. In response to epidermal growth factor, nerve growth factor, and platelet-derived growth factor, B-Raf and Raf-1 are activated, phosphorylate recombinant kinase-inactive MEK-1, and activate wild-type MEK-1. MEK-1 is the dual-specificity protein kinase that selectively phosphorylates MAP kinase on tyrosine and threonine, resulting in MAP kinase activation. B-Raf and Raf-1 are growth factor-regulated Raf family members which regulate MEK-1 and MAP kinase activity in PC12 cells. Protein kinase A activation in response to elevated cyclic AMP (cAMP) levels inhibited B-Raf and Raf-1 stimulation in response to growth factors. Ras.GTP loading in response to epidermal growth factor, nerve growth factor, or platelet-derived growth factor was unaffected by protein kinase A activation. Even though elevated cAMP levels inhibited Raf activation, the growth factor activation of MEK-1 and MAP kinase was unaffected in PC12 cells. The results demonstrate that tyrosine kinase receptor activation of MEK-1 and MAP kinase in PC12 cells is regulated by B-Raf and Raf-1, whose activation is inhibited by protein kinase A, and MEK activators, whose activation is independent of cAMP regulation.

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The activation of the chymotrypsin-like activity of the proteasome is regulated by soluble adenyl cyclase/cAMP/protein kinase A pathway and required for human sperm capacitation

MHR Basic science of reproductive medicine ◽

10.1093/molehr/gaz037 ◽

2019 ◽

Vol 25 (10) ◽

pp. 587-600 ◽

Cited By ~ 2

Author(s):

Héctor Zapata-Carmona ◽

Lina Barón ◽

Lidia M Zuñiga ◽

Emilce Silvina Díaz ◽

Milene Kong ◽

...

Keyword(s):

Protein Kinase ◽

Protein Kinase A ◽

Current Knowledge ◽

Human Sperm ◽

Adenyl Cyclase ◽

Sperm Capacitation ◽

Time Treatment ◽

Proteasome Subunits ◽

Pka Pathway ◽

Kinase A

Abstract One of the first events of mammalian sperm capacitation is the activation of the soluble adenyl cyclase/cAMP/protein kinase A (SACY/cAMP/PKA) pathway. Here, we evaluated whether the increase in PKA activity at the onset of human sperm capacitation is responsible for the activation of the sperm proteasome and whether this activation is required for capacitation progress. Viable human sperm were incubated with inhibitors of the SACY/cAMP/PKA pathway. The chymotrypsin-like activity of the sperm proteasome was evaluated using a fluorogenic substrate. Sperm capacitation status was evaluated using the chlortetracycline assay and tyrosine phosphorylation. To determine whether proteasomal subunits were phosphorylated by PKA, the proteasome was immunoprecipitated and tested on a western blot using an antibody against phosphorylated PKA substrates. Immunofluorescence microscopy analysis and co-immunoprecipitation (IPP) were used to investigate an association between the catalytic subunit alpha of PKA (PKA-Cα) and the proteasome. The chymotrypsin-like activity of the sperm proteasome significantly increased after 5 min of capacitation (P < 0.001) and remained high for the remaining incubation time. Treatment with H89, KT5720 or KH7 significantly decreased the chymotrypsin-like activity of the proteasome (P < 0.001). IPP experiments indicated that PKA inhibition significantly modified phosphorylation of proteasome subunits. In addition, PKA-Cα colocalized with the proteasome in the equatorial segment and in the connecting piece, and co-immunoprecipitated with the proteasome. This is the first demonstration of sperm proteasome activity being directly regulated by SACY/PKA-Cα. This novel discovery extends our current knowledge of sperm physiology and may be used to manage sperm capacitation during assisted reproductive technology procedures.

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